This protocol provides a new approach to quantifying branch dynamics of the dendritic arbor and developing neurons using live time lapse imaging and a 3D image annotation software. This technique images and tracks the branch terminals providing a
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Here, we describe the method we employed to image highly motile dendritic filopodia in a live preparation of the Drosophila larval brain, and the protocol we developed to quantify time-lapse 3D imaging datasets for quantitative assessments of dendrite dynamics in developing neurons.